Title Citotoksičnost fumonizina B1 i B2 na A549 i HepG2 stanicama Title (english) Cytotoxicity of fumonisin B1 and B2 on A549 i HepG2 cells Author Ana Lovrinčević Mentor Maja Šegvić Klarić (mentor)Committee member Daniela Jakšić (predsjednik povjerenstva)Committee member Ana-Marija Domijan (član povjerenstva)Committee member Dubravka Vitali Čepo (član povjerenstva)Granter University of Zagreb Defense date and country 2019-09-25, Croatia Scientific / art field, BIOMEDICINE AND HEALTHCARE Abstract Izvanstanični metaboliti plijesni, koji su toksični ili imaju druge negativne biološke učinke na ljude i životinje, čine skupinu mikotoksina. Ti su spojevi vrlo različitih kemijskih struktura te onečišćuju žitarice i druge namirnice, posebice u tropskim krajevima jer viša temperatura i vlažnost pogoduju nastanku i rastu plijesni. Čovjek je najčešće izložen mikotoksinima konzumacijom hrane, no, u nekim slučajevima, može ih i udahnuti, oni mogu proći kroz kožu ili je izloženost parenteralna.
Fumonizini su mikotoksini koje najčešće proizvode vrste F. verticillioides i F. proliferatum. Od svog otkrića, fumonizini se smatraju mikotoksinima uglavnom proizvedenima Fusarium vrstama. B-skupina fumonizina (FB1, FB2 i FB3) se često otkriva u kukuruzu i u hrani na bazi kukuruza. Danas je poznato najmanje 28 različitih fumonizina, a većina njih označena je kao A-, B-, C- i P-serija. Najznačajniji su fumonizini B-serije, a od njih fumonizin B1 (FB1) te fumonizin B2 (FB2) koji su gotovo jednake toksičnosti. U prirodi se FB1 pojavljuje 3 puta češće od FB2. Cilj ovog rada je procijeniti citotoksične koncentracije pri kojoj je preživljavanje stanica smanjeno za 50% (IC50) fumonizina B1 i fumonizina B2 na dvije različite stanične linije: ljudske stanice adenokarcinoma pluća, A549, i ljudske stanice hepatocelularnog karcinoma, HepG2. Određivanje IC50 služi za odabir subcitotoksičnih koncentracija tih dvaju vrsta fumonizina koje će se koristiti u ispitivanju citotoksičnog učinka FB1 i FB2 u kombinaciji s drugim mikotoksinima koji se pojavljuju u kućnoj prašini vlažnih prostora, zraku i u hrani. Kako bi se ispitao citotoksični učinak FB1 i FB2, A549 i HepG2 stanice tretirane su toksinima u koncentracijama od 10 do 500 μM tijekom 24 h. Vijabilnost je određena MTS proliferacijskim testom (490 nm). Na ljudske stanice A549 i HepG2 primijenio se MTS proliferacijski test.
FB1 je slabo citotoksičan za A549 stanice, tj. značajno smanjuje vijabilnost stanica samo nakon primjene u najvećim koncentracijama (400 μM i 500 μM) u odnosu na kontrolu. FB2 povećanjem koncentracije potiče proliferaciju A549 stanica. Stanice HepG2 su osjetljivije na djelovanje oba fumonizina pri čemu FB2 ima nešto jači citotoksični učinak od FB1; FB2 značajno smanjuje vijabilnost stanica u koncentraciji od 150 μM, a FB1 u dvostruko većoj. Za oba fumonizina nije bilo moguće odrediti IC50 u stanicama A549 i HepG2 jer fumonizini, aplicirani u rasponu koncentracija od 10 μM do 500 μM, nisu uzrokovali pad vijabilnosti za više od 50% stanica.
Abstract (english) Extracellular mold metabolites, which are toxic or have other adverse biological effects on humans and animals, form a group of mycotoxins. These compounds are of very different chemical structures and contaminate cereals and other foods, especially in the tropics because high temperatures and humidity favor mold formation and growth. People are often exposed to mycotoxins by consuming food, but in some cases it can be inhaled, they can pass through the skin or the exposure is parenteral.
Fumonisins are mycotoxins most commonly produced by F. verticillioides and F. proliferatum. Since its discovery, fumonisins have been considered mycotoxins mainly produced by Fusarium species. The B-group of fumonisins (FB1, FB2, and FB3) is often detected in maize and maize-based foods. Today, at least 28 different forms of fumonisin are known, and most of them are designated as A-, B-, C-, and P-series. The most significant are B-series fumonisins, of which fumonisin B1 (FB1) and fumonisin B2 (FB2) are of almost equal toxicity. In nature, FB1 occurs 3 times more frequently than FB2.
The aim of this study is to evaluate the cytotoxic concentrations at which cell survival is reduced by 50% (IC50) of fumonisin B1 and fumonisin B2 on two different cell lines: human lung adenocarcinoma cells, A549, and human hepatocellular carcinoma cells, HepG2. The determination of IC50 is used to select the subcytotoxic concentrations of these two types of fumonisins to be used in the study of the cytotoxic effect of FB1 and FB2 in combination with other mycotoxins occurring in dust of humid, air and food. To examine the cytotoxic effect of FB1 and FB2, A549 and HepG2 cells were treated with toxins at concentrations of 10 to 500 μM within 24 h. The viability was determined by MTS proliferation assay (490 nm). MTS proliferation assay was applied to human A549 and HepG2 cells.
FB1 is poorly cytotoxic to A549 cells, that means that it significantly reduces cell viability only after administration at the highest concentrations (400 μM and 500 μM) relative to control. FB2 promotes proliferation of A549 cells by increasing concentration. HepG2 cells are more sensitive to the action of both fumonisins with FB2 having a slightly stronger cytotoxic effect than FB1; FB2 significantly reduces cell viability at a concentration of 150 μM and FB1 at twice as high. For both fumonisins it was not possible to determine IC50 in A549 and HepG2 cells, because the fumonisins, administered in the concentration range of 10 μM to 500 μM, didn`t cause a viability decrease of more than 50% of the cells.
Keywords
mikotoksini
fumonizin B1
fumonizin B2
plućne stanice
jetrene stanice
citotoksičnost
Keywords (english)
mycotoxins
fumonisin B1
fumonisin B2
lung cells
liver cells
cytotoxicity
Language croatian URN:NBN urn:nbn:hr:163:713637 Study programme Title: Pharmacy Type of resource Text File origin Born digital Access conditions Open access Terms of use Created on 2021-04-28 15:41:24
